Bioorthogonal Copper Free Click Chemistry for Labeling and Tracking of Chondrocytes In Vivo
- Bioorthogonal Copper Free Click Chemistry for Labeling and Tracking of Chondrocytes In Vivo
- 권익찬; 김광명; 류주희; 이상민; 장혜윤; 윤화인; 이지영; 나진희; 이혁진; 강선웅; 이슬기; 조영우
- Bioorthogonal Click Chemistry; DBCO-650; Cell therapy; Chondrocytes
- Issue Date
- Bioconjugate chemistry
- VOL 27, NO 4-936
- Establishment of an appropriate cell labeling and tracking method is essential for the development of cell-based therapeutic strategies. Here, we are introducing a new method for cell labeling and tracking by combining metabolic gylcoengineering and bioorthogonal copper-free Click chemistry. First, chondrocytes were treated with tetraacetylated N-azidoacetyl-D-mannosamine (Ac(4)ManNAz) to generate unnatural azide groups (-N-3) on the surface of the cells. Subsequently, the unnatural azide groups on the cell surface were specifically conjugated with near-infrared fluorescent (NIRF) dye-tagged dibenzyl cyclooctyne (DBCO-650) through bioorthogonal copper-free Click chemistry. Importantly, DBCO-650-labeled chondrocytes presented strong NIRF signals with relatively low cytotoxicity and the amounts of azide groups and DBCO-650 could be easily controlled by feeding different amounts of Ac4ManNAz and DBCO-650 to the cell culture system. For the in vivo cell tracking, DBCO-650-labeled chondrocytes (1 x 10(6) cells) seeded on the 3D scaffold were subcutaneously implanted into mice and the transplanted DBCO-650-labeled chondrocytes could be effectively tracked in the prolonged time period of 4 weeks using NIRF imaging technology. Furthermore, this new cell labeling and tracking technology had minimal effect on cartilage formation in vivo.
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